SPECIFIC PRIMERS FOR DETECTION OF ALICYCLOBACILLUS-ACIDOTERRESTRIS BYRT-PCR

The reverse transcription-polymerase chain reaction (RT-PCR), after a short enrichment culture, was used to detect Alicyclobacillus acidoter restris in acidic beverages. Two specific primers were selected from t he region of V2 and V4 on 16S rRNA gene. With this primer set, 294-bp fragments from ii. acidoterrestris could be amplified. The detection l imit of the RT-PCR with the FHLP filters was about 10(-1) fg of pure t otal RNA per reaction. Juice samples inoculated with 10(4) cfu of A. a cidoterrestris per mi were RT-PCR positive without enrichment. However , after 15 h of enrichment, the samples inoculated with 2-3 cfu ml(-1) were positive. This RT-PCR culture assay would enable rapid and speci fic detection of strains of A. acidoterrestris in acidic beverages.